DNA sequencing by parallel olgonucleotide extensions

Abstract

Disclosed are oligonucleotides probes which are useful for determining a sequence of nucleotides in a target ploynucleotide. In one embodiment, the probe is of the formula: HO-(3')(B)j(5')-OP(═O-)NH-(B)k-Bt*, wherein B is a nucleotide or an analog thereof; j is in range of from 1 to 12; is in the range of from 0 to 12, such that the sum of j and k is less than or equal to 12; and Bt* is a labeled chain-terminating moiety. In s second embodiment, the probe has a formula selected from the group consisting of (1) OP(═O)(O--)O--(5═)(B)sRRRR(B)wBt*, and (2) OP(═O)(O--)(5')(B)sRRRR(B)w(3')OP(═O)(O--)O, wherein: is a deoxyribonucleotide or an analog thereof; R is a ribonucleotide; s is in the range of from 1 to 8; w is in the range of from 0 to 8, such that the sum of j and k is less than or equal to 8; and Bt* is a labeled chain-terminating moiety.

I claim: 
     
       1. An oligonucleotide probe of the formula:   HO--(3')(B)j(5')--OP(═O)(O--)NH--(B)k-Bt*     wherein:   B is a nucleotide or an analog thereof;   j is in the range of from 1 to 12;   k is in the range of from 1 to 12, such that the sum of j and k is less than or equal to 12;   Bt* is a labeled, non-extendable chain-terminating moiety.   
     
     
       2. An oligonucleotide probe selected from the group consisting of:   OP(═O)(O)O--(5')(B)sRRRR(B)wBt*     and     OP(═O)(O--)O--(5')(B)sRRRR(B)w(3')OP(═O)(O--)O     wherein:   B is a deoxyribonucleotide or an analog thereof;   R is a ribonucleotide;   s is in the range of from 1 to 8;   w is in the range of from 0 to 8, such that the sum of s and w is less than or equal to 8; and   Bt* is a labeled, non-extendable chain-terminating moiety.